Evaluation of wheatstraw, sawdust, banana fronds, maize cobs and cotton hulls substrate combinations for Pleurotus ostreatus cultivation.

Evaluation of wheatstraw, sawdust, banana fronds, maize cobs and cotton hulls substrate combinations for Pleurotus ostreatus cultivation. | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Evaluation of wheatstraw, sawdust, banana fronds, maize cobs and cotton hulls substrate combinations for Pleurotus ostreatus cultivation. Silivani Elyson, ChisangoTawanda Jonathan, Chitindingu Kudakwashe, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4831136/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The yield and quality of oyster mushroom is largely dependent on the chemical and nutritional content of the substrate. The objective of the study was to determine the effect of different individual substrates and substrate combinations on the perfomance of P.ostreatus for growth and sustainable development of the mushroom cultivation industry in Zimbabwe. Various individual substrates used as treatments, selected for the cultivation of oyster mushrooms were, wheatstraw, cotton hulls, sawdust, maize cobs and banana fronds each of 1kg and replicated 4 times. Various substrate combinations, also used as treatments were, cotton hulls and wheatstraw, cotton hulls and sawdust, cotton hulls and maizecobs, cotton hulls and banana fronds in the ratio 1:1. Each combination weighed 1kg and was replicated 4 times.Compound substrates perfomed much better in terms of biological efficiency and spawn run compared to individual substrates. The highest biological efficiency (76%) and spawn run(17 days) were obtained from combining cotton hulls and sawdust. Spawn run was fastest(12 days) for cotton hulls and wheatstraw combination. Analysis in terms of economic return, revealed that mushroom production was most profitable using cotton hulls and sawdust as substrate with a benefit-cost ratio of 5.7 compared to other combinations of agricultural residues. In terms of spawn run, yield and economic return, combining cotton hulls and sawdust is highly recommended for farmers involved in the cultivation of Pleurotus ostreatus to satisfy increasing consumer demand, whilst decreasing pressure on limited natural resources and preventing ecosystems degradation. Substrate combination mushrooms Pleurotus ostreatus biological efficiency spawn run cost-benefit ratio Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Pleurotus ostreatus( oyster) is a member of Pleurotus species of mushrooms cultivated globally for food. According to Besufekud et al (2019), oyster mushrooms are the second largest commercially produced mushrooms in the world. They possess a rare capability to convert lignocellulosic waste residues into edible fruiting bodies, rich in nutrients. Oyster mushrooms are reported by Ramsbottom ( 1954 ) to contain high amounts of vitamins, particularly vitamin D and high protein contents because they contain enzymatic proteases that can breakdown complex macromolecules in agricultural wastes into individual amino acids. Of much importance is the absence of fatty acids in oyster mushrooms, which makes them ideal for persons willing to maintain a low fat diet. Furthermore, oyster mushrooms contain the appropriate ratios of potassium and sodium, which are essential in reducing heart disease. In Zimbabwe, mushroom cultivation is still at the grassroots level with most small-scale farmers producing enough for subsistence. Much knowledge is still lacking pertaining to the utilisation of substrate individually and in combination to optimise production and achieve commercial production leading to exportation.Govera (2020) Mushroom substrate is the material or substance, which provides food for the growing mycelium. Oyster mushrooms can grow on a wide range of substrates compared to other mushrooms. They can grow on all wood types including sawdust and paper pulp sludge. They also grow on all types of cereal straw such as maize straw, wheatstraw and rice straw. They also grow well on other types of agricultural substrates such as corncobs, banana fronds, maize stalks and leaves, savanna veld grass, sage grass, cottonseed hulls and coffee grounds. The mechanism by which substrate is broken down, is by the extracellular release of enzymes.According to Onyeka et al ( 2017 ) the extracellular enzymes secreted by fungi contain amorphous homo and heteropolysaccharides often associated with fungal protein. These enzymes bind to cellulose, lignin and hemicellulose, and break them down into simple sugars and peptides. Sayner et al (2019) alludes that the yield and quality of oyster mushroom is largely dependent on the chemical and nutritional content of the substrate. In instances where, a nutrient lacking in one substrate it is then provided by another substrate. There is need to establish a consensus by, pulling high yielding substrates together and then come up with the best formulation. Mushroom yield is closely associated with the chemical and biological composition of the substrate (Bhattacharjya et al 2015 ) Most mushroom farmers in Zimbabwe today are using cotton hulls to grow oyster mushroom. As more people are becoming knowledgeable about cotton hulls as substrate, it creates a demand for this commodity. Tapiwa (2021). As a result of the increase in demand for cotton hulls there is increase in price of cotton hulls. It becomes economically non profitable to venture into mushroom farming due to expensive substrate. The price of wheat straw and cotton hulls is increasing every year, as many more people become mushroom farmers. This in turn affects the pricing of the mushrooms themselves. Dead plant matter has different composition of nutrition. There are many factors, which determine selection of a particular substrate. These are nutrition, cost, availability, accessibility and storage. A single particular substrate may be readily available but costly to the farmer. The length of the mushroom crop or the number of flushes depends on the substrate used for the cultivation of oyster mushrooms. For example, wheatstraw has good nutrition but it has a limited cropping life. Banana fronds also have good nutrition but they has low water retention. Sawdust has high nutrition but it takes longer to fruit. Cotton hulls are the best for fruiting, spawn run, have good nutrition but they are very expensive and seasonal (Pant et al 2006 ). There are limitations in nutritional composition when only one substrate is used to grow mushrooms. A formulation of substrates might contribute to better nutrition in oyster mushrooms. The best composition of substrates should have a short spawn run, a high biological efficiency and a high benefit cost ratio. C/N ratio directly affects the mushroom spawn run, yield and biological efficiency. Spawn run Spawn run is the time it takes for the substrate to be fully covered by the mushroom mycelia. It is affected by qualities of the substrate and qualities of the spawn. For a shorter spawn run a substrate should have a high surface area and good nutrition. Banana leaves require 19 days for spawn run.(Asmamaw et al 2014).Sawdust takes 30 days for spawn run. (Hoa et al 2015).Cotton hulls takes 18 days for spawn run.(Muswati et al 2021). Wheatstraw takes 16 days for spawn run.(Girmay et al 2016 ). Corn cobs takes 40 days for spawn run. (Itelima 2011 ) Biological efficiency Biological efficiency is defined as the percentage measurement of harvested fresh mushrooms relative to the dry weight of substrate.(Chukwurah et al 2012). In theory a 100% biological efficiency is achieved when fresh mushrooms having a moisture content of 90% are havested from a substrate with a moisture content of 75% of which 25% of dry substrate gives us the mushrooms. BE = DP/M BE-Biological Efficiency DP-mass of fresh mushrooms produced in kgs M-initial dry mass of substrate in kgs In practice a 100% biological efficiency is achieved when a mushroom farmer collects 10 kgs of mushrooms from three harvests after cultivating on 10kgs of dry substrate. Outside factors such as temperature and humidity, affect biological efficiency. (Girmay 2016) A biological efficiency above 75% will be economical for the mushroom farmer. This is only achievable by using a high spawn rate. Usually the first, second and third flushes are the best flushes, with each flush decreasing in yield form the first. For subsistence, small scale farmers can allow the mushroom bags to fruit until the substrate is spent. For commercial farming only three flushes are economical. (Zakil et al 2019) B-C Ratio B-C (Benefit-Cost) ratio is the ratio of benefit from selling output monetary terms relative to the total cost during production. However, as the value of B:C increases so does the feasibility of mushroom production in economical aspect. To find the ratio the yield of mushrooms by is divided by the the sum total expenditure of raw materials and substrate. Rice straw gives a B-C ratio of 3.498, wheatstraw 1.108, sugarcane bargasse 0.217 and banana leaves 1.408. Therefore, as the B-C ratio becomes greater then 1, the more profitable the project and as the B-C ratio falls below 1, the less profitable the project. (Dubey et al 2019 ). Carbon to Nitrogen ratio According to Kashangura et al ( 2005 )for mycelial growth and fruiting body formation the carbon to nitrogen ratio should be 50: 1, 100: 1 or 500: 1. Most agricultural wastes such as corncob, grain straws, wood, sawdust and banana fronds have sufficient C; N ratios to support saprophytic mushroom growth. C: N ratio is the proportion of carbon to nitrogen in a substrate. (Hoa et al 2015) Oyster mushroom mycelial growth is optimum in the presence of more carbon compared to nitrogen. Therefore, substrates comprised primarily of cellulose, hemicellulose and lignin are good candidates for oyster mushroom cultivation. In this study, we will investigate cotton hulls, sawdust, wheatstraw, banana fronds and maize cobs individually and in combination to determine potential in optimisation of Pleurotus ostreatus . We discuss the characteristic features and C: N ratios of the substrates under investigation below: Banana fronds are the dried leaves from banana plants. During the dry season banana leaves lose a lot of moisture and as a result, the plant withholds water in the stem, leading to the lower leaves turning brown and then drooping down the plant. Banana farmers have no choice but to cut away the leaves and burn them because the leaves invite pests such as mosquitoes, rats and fungi, which feed on the dead leaves. The carbon to nitrogen ratio in banana leaves is 70:1. (ScienceMate 2022 ) Wheatstraw is the grass or straw that farmers leave behind when wheat farmers harvest their wheat. Wheat is a winter crop with a very short season and farmers have to use combine harvesters to harvest the wheat before the pod breaks and disperses the seed. Most farmers use a bailer to collect the leftover straw and then store it for feeding their livestock. Wheatstraw has problems with storage since it easily burns and termites tend to feed on the wheat straw. Less knowledgeable farmers tend to burn the remaining wheatstraw left in the fields in preparation for the maize farming season in Zimbabwe. This however leads to veld fires and deforestation. The carbon to nitrogen ratio in wheat is 80:1. (Lentz and Lindsey 2016) Maize cobs are the cob that remains after removal of maize seeds from the cob. Large and huge masses of this waste accumulates soon after maize has been harvested because fresh maize cobs are sold and cooked as breakfast and discarded along streets and at dumpsites creating a huge environmental problem. Most of the weight of the maize cob is in the cob itself and this proves that it has a lot of carbon. Grinding down the cob after drying it can drastically increase its surface area thereby allowing mushroom mycelia to spread and absorb nutrients from the cob. Postharvest, farmers discard the cobs after shelling and most small-scale farmers use them as firewood during the winter. The carbon to nitogen ratio of maize cobs is 100:1.Adjapong (2015) The wood industry is reponsible for producing furniture and roofing material. However, during the cutting down, shaping and designing a lot of this wood is broken down into wood shavings and sawdust. However if the wood waste derived from this industry was to be saved stored and commercialised for mushroom farming we could archive food security. The carbon to nitrogen ratio of wood is in the range of 500:1. (Rynk 1992 ) Cottonseed hulls are a waste product resulting from the pressing and crushing down of cottonseed into cottonseed cooking oil. Ginneries opt to burn or throw away the waste as the main method of disposal. However, cottonseed hulls have proved to contain high amounts of nitrogen and carbon essential for mushroom growth. The carbon to nitrogen ratio of cottonseed hulls is 59:1. (Haziran 2019 ) This study will provide sufficient data to make decisions on the utilisation of different types of agricultural and forest waste into nutritional food. To analyse, evaluate and ultimately identify the most appropriate composition of substrate for cultivating Pleurotus ostreatus . Materials and methods Site of the study The research was conducted in a steel Mushroom House, at the Department of Biotechnology, School of Health Sciences and Technology, Chinhoyi University of Technology, Zimbabwe from January to March 2022 for evaluating the effect of different substrates on the perfomance of Pleurotus ostreatus . Treatments The substrates used in this investigation were wheatstraw, cottonseed hulls, sawdust, banana fronds and maize cobs. Each substrate combination weighing 1kg (dry weight) at a ratio of 1:1 was also distributed into a single factor experiment laid out in a Completely Randomised Design Design. Nine treatments were replicated four times thus making 36 bags. Analysis of variance tested for differences among means and means separated using Least Significant Difference (LSD) at the 5% level of significance. One-way analysis of variance (ANOVA) with Duncan’s multiple range tested the mean significant differences ( p < 0.05) among treatments by using computer software R Statistical Package. Substrate collection and bag filling: Different substrates were collected from different locations. Cotton hulls purchased from a retailer in Harare. Maize cobs purchased from Citrus farm in Chinhoyi. Banana fronds collected from Bhagudha farm Chinhoyi. Sawdust procured from Chinhoyi University Works and Estates Department. Wheatstraw was collected from Chinhoyi University farm Chinhoyi. Maize cobs, wheatstraw and banana fronds were ground at the Chinhoyi University feed factory. Maize cobs were ground into powder, wheatstraw into chunks 2-4cm and banana fronds into chunks 2–4 cm long. Combinations formulated with respect to cotton hulls at a ratio of 1:1 dry weight. They were soaked with clean tap water to remove all dust particles. Then the substrate combinations filled into separate 200litre drums and a mixture of 4% sodium hypochlorite and 20% cacium hydroxide solution added to cover the substrate. The drums were covered and left overnight. The following day, substrates were drained of excess water separately on steel fence tables. Each substrate was mixed with an equal proportion of cotton hulls. Inoculation/spawning: We made our own Pleurotus ostreatus grain spawn to avoid bias. Spawning was carried out in transparent polythene pastic bags in layers subsequently with the first layer consisting of substrate and the top layer covered with spawn. Each bag comprised of 3 layers of substrate and 3 layers of spawn. After inoculation each bag approximately measured 2kg of substrate combinations by wet weight. Then the mouth of each bag was secured tightly with string and small holes were punched on the top and lateral sides of each bag for aeration. Incubation: The packed bags were incubated in a dark steel mushroom house until the mycelium penetrated to the bottom of the bag. The bags were hung from trusses welded below the roof of the mushroom house to allow excess water to drip from the bottom of the bag, to prevent direct contact of the bags with the floor of the mushroom house and to prevent contact with insects such as ants and termites. Growing: From the moment bags were introduced into the mushroom house the spawn run was observed and measured until the whole bags were covered with white mycelium. Thereafter the plastics were removed from the bags and the bags were placed on a wooden platform 30cm from the floor with a space of 20cm inbetween bags. When pinheads started appearing a light source was introduced into the mushroom house at night and watering commenced 3 times a day at 6 hour intervals using a knapsack sprayer. Harvesting: After 5 days of watering mushrooms were harvested and appropriate observations and measurements were taken. Harvesting commenced when the cap attained the maximum diameter, just before the edges flatten out. Firstly the whole bag and mushroom weight was recorded, then picking was done by gently twisting and pulling out the stalk. Mushrooms were then weighed and packaged into 200g punnets, wrapped with transparent plastic wrapper and sold for $ 1/200g. Then the weight of the empty bag was recorded. The benefit from selling the mushrooms was also recorded. Data collection and analysis: Data was recorded periodically during the incubation from colonisation to final substrate weight. Parameteres under conisderation were number of days to full colonisation, weight of bag and mushroom, total mushroom yield, final substrate weight, total mushrooms sold and benefit cost ratio. Collected data were analysed using R statistical software and Microsoft Excel 2007. Analysis of variance (ANOVA) was used to test among treatments and means were separated using Least Significance Difference (LSD) at the 5% level of significace. Results and Discussion The yield and yield attributing observations of mushrooms obtained from different combinations of substrates were compared. There was notable and considerable variation with respect to spawn run, quantiy of first yield and final substrate weight. Effect of substrate combinatins on spawn run and 1st yield weight: Highly significant results were observed among treatments in terms of spawn run of Pleurotus ostreatus as shown in Fig. 3 . Of all the combinations under investigation wheatstraw and cotton hulls combination took the shortest time (12 days) followed by cotton hulls and banana fronds(13 days), cotton hulls and maizecobs(13 days) and lastly cotton hulls and sawdust(17 days). Individual substrate took longer to complete spawn run compared to combinations i.e wheatstraw(16 days), banana fronds(17 days), maize cobs (19 days), cotton hulls(19 days ) and finally sawdust(20 days). This coincides with the findings of Muswati et al 2021 that combinations of cotton hulls perform much better during spawn run compared to individual substrates. Table 1 Mean values of first yield, spawn run and final substrate Substrate First yield(g) Spawn run(days) Substrate final weight(g) Cotton hulls and wheatstraw 376 12 1576 Cotton hulls and banana fronds 449 13 1716 Cotton hulls and sawdust 428 17 1099 Cotton hulls and maize cobs 420 13 1508 Cotton hulls 400 19 1213 Wheatstraw 212 16 980 Sawdust 155 20 1417 Banana fronds 250 17 1118 Maizecobs 263 18 1842 Anova table for analysis of variance on substrate means Df Sum Sq Mean Sq F value Pr(> F) tm 3 11217444 3739148 146.8 < 2e-16 *** Residuals 32 815253 25477 --- Signif. codes: 0 ‘***’ 0.001 ‘**’ 0.01 ‘*’ 0.05 ‘.’ 0.1 ‘ ’ 1 At the 5% level of significance a P value of < 2e-16 is less than 0.05 which means there are significant differences among treatment means. Effect of different substrates on total yield,spawn run and final substrate weight of oyster mushroom. ( Table 1 ). The weight of substrate combination after first harvest was lowest in case of cotton hulls and sawdust(1099g) followed by cottonhulls maizecobs, cottonhulls-wheatstraw and cottonhulls-banana fronds at 1508g, 1576g and 1716g respectively as shown in Table 1 . This is evidence that oyster mushroom absorbs nutrients more efficiently in substrates composed of cotton hulls and sawdust compared to other substrate combinations of cotton hulls. This might also give an indication of particle size and absorption efficiency given the small particle size of sawdust rendering nutrient accessibility easier. Thus, it also indicates that the high biological efficiency obtained from cottonhulls-sawdust combination (76%) is attributed to the small surface area. Statistical analysis of data generated showed that there are significant differences in the total yield of mushrooms obtained from different substrate combinations. Yields from individual substrates pale in comparison to substrate combinations with the exception of cotton hulls(400g) alone which exceeded combinations of cotton hulls and wheatstraw(376g). Combinations of substrates contribute to nutrient compensation as what is lacking in one substrate is supplemented from the other. The high biological efficiency exhibited by cotton hulls sawdust combination may be attributed to high lignolytic and cellulonitic activity of the substrates. Muswati et al (2021)cultivated Pleurotus ostreatus on cotton hulls and baobab fruit shells and observed similar results. Economic analysis(Table 2 ) Cost of cultivation: It is analysed based on the cost of different materials and combinations of substrate. However, in this research the cost of required materials was at a constant for each treatment, except for cost of substrates, which are shown in Table 2 : B-C ratio: The B: C ratio relates the benefit from selling mushrooms in monetary terms to the total cost during production. In this case it was calculated in United States Dollars by dividing the cost of production per kilogram of substrate by the total benefit of selling fresh mushrooms per kilogram of substrate. Cost of producton entails total expenses of raw materials. Benefit of production is calculated by dividing the biological efficiency by the price of selling mushrooms per kilogram of substrate. Substrate combinations were considered profitable if the B: C ratio was greater than 1. The differences in B:C ratio are shown in the Table 2 . The highest most significant ratio was produced by combination of cotton hulls and sawdust(5.7) whereas the lowest least significant ratio was produced by combination of cotton hulls and wheatstraw. For individual substrates cotton hulls produced a significant ratio of 4.24 which was more profitable compared to combinations of cotton hulls with banana fronds (2.43), cotton hulls with wheatstraw (2.02) and cotton hulls with maizecobs(3.73). For individual substrates cotton hulls(4.24) were more profitable compared to all the other substrates with the least profitable being banana fronds with a B;C ratio of 1.28. The data generated here is empirical basis for encouraging mushroom farmers to consider combining cotton hulls and sawdust for achieving high economic return with low investment. We suggest the low yied from wheatstraw, maizecobs, sawdust and banana fronds emanating from their high water retenton tendencies in contrast to cotton hulls. Furthermore wheatstraw and banana fronds lack compactness due to the low surface area which impedes complete ramification of substrate. Combinations of cotton hulls with wheatstraw and banana fronds did not improve yield due to reasons previously stated. Sawdust and cotton hulls proved to be the ideal combination firstly as a result of the large surface area then secondly due to the rich ratios of lignin and cellulose from sawdust combined with nitrogen and xylanases provided by cotton hulls.(Vhargese and Amritkumar 2020) Table 2 Mean values of B:C ratio of individual substrates and combinations Substrate Biological efficiency/kg Cost/kg(US $ ) Benefit(US $ )/kg B:C ratio Cotton hulls and wheatstraw 34 0.84 1.7 2.02 Cotton hulls and banana fronds 45.6 0.94 2.28 2.43 Cotton hulls and sawdust 76.3 0.67 3.82 5.7 Cotton hulls and maize cobs 50 0.67 2.5 3.73 Cotton hulls 60.3 0.71 3.015 4.24 Wheatstraw 25.2 0.68 1.26 1.85 Sawdust 18.5 0.51 0.925 1.81 Banana fronds 20 0.78 1 1.28 Maizecobs 26.3 0.57 1.315 2.31 Spawn run was fastest for wheatstraw(16days) due to high nutrient availability and aeration. As the degreee of aeration decreases as a result of compactnaess of the substrate there is increased longevity of the spawn run as observed for sawdust(20 days) as shown in Fig. 1 .. Spawn runner was also faster for banana fronds (17 days) also due to availability of aeration. Maizecobs and cotton husks managed to achieve the same rate of spawn runner dur tocompactness. It was observed that irregardless of nutrient availability particle size of substrate directly affects availability of air pockets within the substrate thereby limiting aiflow and affecting rate of fermentation. The more compact a substrate is the lesser the airation results in a slow spawn run. Biological efficiency is directly affected by the carbonn to nitrogen ratio. The more nitrogen there is in a substrate the higher the rate of protein synthesis leading to a higher yield of mushrooms(cotton husks). In contrast the more carbon there is in a substrate the more the more enegy available for cellular replication(mitosis) but less energy available for protein sythesis,as in the case of wheatstraw and sawdust. However complentation comes into play when the shortfalls of one substrate is covered by suplementation with another substrate resulting in a high biological efficiency as in the case of cotton hulls and sawdust as shown in Fig. 2 . Spawn run is fastest for cotton husk and wheatstaw combinations. Spawn run is slowest for cotton husks and sawdust. The imrovement in spawn run for cotton hulls from 19 days to a shorter time of 12 days is abbtributed to increased aeration. Furthermore there is a notable decrease in the spawn ru for sawdust from 20 days to 17 days attributed by increased carbon to nitrogen ratio and aeration. Overally combinations of substrate perfomed better compared to indiividual substrate across the board. Figure 4 is data translated from Table 2 giving a graphical representation of the benefit cost ratio of using combinations of substrated compared to individual substrate. The highest benefit being afforded by combining cotton hulls and sawdust and the lowest being cotton hulls and wheatstraw. The only exception being cotto husk individully outperforming combinations with maizecobs, wheatstraw and banana fronds. Figure 5 shows appearance of oyster mushrooms cultivated on different combinations of substrate. On cotton hulls and sawdust, oyster mushrooms are bigger and there is uniform distribution of bunches right round the bag, there is fruiting on all sides of the bag. On cotton hulls and banana fronds there are lesser bunches of mushrooms even though there is also an even distribution of mushrooms right round the bag. On cotton houlls and wheatstraw the mushrooms have a much thicker skin and appear more grey than other mushrooms grown on other combinations. However, there are fewer mushrooms only appearing on one side of the bag. On cotton hulls and maizecobs there is a uniform distribution of mushrooms right round the bag however, the mushrooms appear smaller than the ones on cotton hulls and sawdust. The mushrooms on cotton hulls and maizecobs combination are whiter than mushrooms grown on other mushrooms. Conclusion This study was conducted by growing Pleurotus ostreatus on 5 different agricultural waste substrates namely cotton hulls, maizecobs, wheatstraw, banaba fronds, sawdust and on combinations of 4 substrates with respect to cotton hulls. Among the individual substrates wheatstraw colonised the fastest whereas among the combinations wheatstraw and cotton hullls were colonised fastest. In terms of biological efficiency, individually cotton hulls performed exceedingly well compared to other substrates. However, combinations of substrates revealed a combination of sawdust and cotton hulls to be superior to other combinations. Combinations of substrates proved to be highly efficient both in spawn run and biological efficiency compared individual substrates. In terms of economic return, combinations of sawdust and cottonhulls (5.7) proved to be the most profitable. Hence, we conclude that in terms of spawn run, yield and economic return combination of cotton hulls and sawdust is highly recommended for farmers involved in the cultivation of Pleurotus ostreatus . Declarations Ethical approval Since no humans or animals were involved in this study no ethical committees were consulted. Consent to participate is not applicable since no human or animal participants were required. All supervisors and fellow co-authors have provided their consent to publish. Funding This study was self funded by the corresponding author Elyson Silivani as an Mphil Student in the School of Health Sciences, Department of Biotechnology, Chinhoyi University of Technology. 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Sintim,(2015) Maize Residue as a Viable Substrate for Farm Scale Cultivationmof Oyster Mushroom (Pleurotus ostreatus) , Hindawi Publishing Corporation Advances in Agriculture Volume 2015, Article ID 213251, http://dx.doi.org/10.1155/2015/213251 Rynk, R., ed. 1992. On-Farm Composting Handbook . Northeast Regional Agricultural Engineering Service.. Available on-line at : http://www.cals.cornell.edu/dept/compost/OnFarmHandbook/apa.taba1.html , or in print from : Northeast Regional Agricultural Engineering Service, 152 Riley-Robb Hall, Cornell University, Ithaca NY 14853. Haziran,(2019), Cottonseed hulls:Oyster mushroom cultivation , available online at https://www.kulturkia.com/2019/06/cottonseed-hulls-mushroom-compost.html Accessed 16/01/2022 Chukwurah, N. F.1, Eze, S. C.2, Chiejina, N. V.3, Onyeonagu, C. C.2*, Ugwuoke, K. I.2, Ugwu, F. S. O.1, Nkwonta, C. G.1, Akobueze, E. U.1, Aruah, C. B.1 and Onwuelughasi,(2012) C. U. Performance of oyster mushroom (Pleurotus ostreatus) in different local agricultural waste materials , African Journal of Biotechnology Vol. 11(37), pp. 8979–8985, DOI: 10.5897/AJB11.2525 ISSN 1684–5315 Available online at http://www.academicjournals.org/AJB Zakil Fathie Ahmad, Sueb Mohd Shafiq Mohd,Isha Ruzinah (2019) Growth and yield performance of Pleurotus ostreatus on various agro-industrial wastes in Malaysia , AIP Conference Proceedings 2155, 020055 https://doi.org/10.1063/1.5125559Besufekud Girmay, Z., Gorems, W., Birhanu, G.(2016).. Growth and yield performance of Pleurotus ostreatus (Jacq. Fr.) Kumm (oyster mushroom) on different substrates . AMB Expr 6, 87 https://doi.org/10.1186/s13568-016-0265-1 Muswati Charles, Simango Kennedy, Tapfumaneyi Linda, Mutetwa Moses, Ngezimana Wonder,(2021) The Effects of Different Substrate Combinations on Growth and Yield of Oyster Mushroom (Pleurotus ostreatus) Hindawi International Journal of Agronomy Volume 2021, Article ID 9962285, Available online at https://doi.org/10.1155/2021/9962285 Varghese Bably Pearl, Amritkumar Pavithra,(2020) Comparative Study on Cultivation of Oyster Mushrooms using Nutrition Enhancing Substrates, International Journal of Scientific Research in Biological Sciences Vol.7, Issue.2, pp.105–111, April (2020) E-ISSN: 2347-7520 DOI: https://doi.org/10.26438/ijsrbs/v7i2.105111 Itelima J. U.,(2011), Cultivation of mushroom (Pleurotus ostreatus) using corn cobs and saw dust as the major substrates, Global journal of agricultural sciences vol 11, no. 1, 2012: 51–56 DOI: http://dx.doi.org/10.4314/gjass.v11i1.9 Additional Declarations No competing interests reported. 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Elyson","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBklEQVRIiWNgGAWjYDACdgjFw8cMogxsgARj4wG8WpihWtggWtJAWhqI0sLABqEOg0m8WviZeYw/fNxRJ8PGzmP4uaDgvN3a9sNAW2psonFpkWzmMZOceeYw0GE8xtIzDG4nbzuTCNRyLC23AYcWg8M8Zsy8bQeAWtgSpHmAWswOALUwNhzGqcX+MI/xZ962OpCW5N88BueSzc4/xK/FgJnHQJq3jRmohfkY0JYDdmY3CNgicZitTHJm22GwFmseg+QEsxtAWxLw+IW/vXnzh49tdfb8/Aebb/P8sbM3O5/+8MGHGhucWhgYOAxQuIlglQk4lYMA+wMUrj1exaNgFIyCUTAiAQBGCFRTFcTtLgAAAABJRU5ErkJggg==","orcid":"","institution":"Chinhoyi University of Technology","correspondingAuthor":true,"prefix":"","firstName":"Silivani","middleName":"","lastName":"Elyson","suffix":""},{"id":345015394,"identity":"8119a8ee-4815-4536-9049-6ef22210b909","order_by":1,"name":"ChisangoTawanda Jonathan","email":"","orcid":"","institution":"Chinhoyi University of Technology","correspondingAuthor":false,"prefix":"","firstName":"ChisangoTawanda","middleName":"","lastName":"Jonathan","suffix":""},{"id":345015396,"identity":"7f480a02-7425-424d-87a9-d8c3f199f586","order_by":2,"name":"Chitindingu Kudakwashe","email":"","orcid":"","institution":"Chinhoyi University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Chitindingu","middleName":"","lastName":"Kudakwashe","suffix":""},{"id":345015397,"identity":"00480be5-ca43-4df4-9016-2e7ef7d0e9bd","order_by":3,"name":"ChiguNomathemba Loice","email":"","orcid":"","institution":"Chinhoyi University of Technology","correspondingAuthor":false,"prefix":"","firstName":"ChiguNomathemba","middleName":"","lastName":"Loice","suffix":""}],"badges":[],"createdAt":"2024-07-30 21:08:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4831136/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4831136/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":63812307,"identity":"2717e570-29db-4da0-b0f7-480618e118eb","added_by":"auto","created_at":"2024-09-02 14:11:03","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":159619,"visible":true,"origin":"","legend":"\u003cp\u003eMean values of \u003cem\u003ePleurotus ostreatus\u003c/em\u003e spawn run expressed over 1 day intervals observed on individual substrates\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4831136/v1/0fe69e862bc546880ffb23ad.png"},{"id":63812309,"identity":"be09c686-f03a-43a1-8f3d-44789a47c92e","added_by":"auto","created_at":"2024-09-02 14:11:03","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":35856,"visible":true,"origin":"","legend":"\u003cp\u003eMean values of Pleurotus ostreatus Biological efficiency observed on individual substrates and on combinations with cotton hulls.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4831136/v1/0f2a012e44ef85e8039f7aa6.png"},{"id":63812308,"identity":"c9b56b6d-1091-4f21-97aa-d7371c57df7d","added_by":"auto","created_at":"2024-09-02 14:11:03","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":228504,"visible":true,"origin":"","legend":"\u003cp\u003eMean values of Pleurotus ostreatus spawn run expressed over 1 day intervals observed on individual substrates and combinations of substrates\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4831136/v1/05521c9cd668fc44f2bc2eb7.png"},{"id":63812306,"identity":"6c6fac14-6bf6-4e1c-a18c-d1db9057c71f","added_by":"auto","created_at":"2024-09-02 14:11:02","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":25864,"visible":true,"origin":"","legend":"\u003cp\u003eCost/benefit ratios Pleurotus ostreatus mushroom cultivated on different combinations of substrates.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4831136/v1/685796333a1825a1b47cf045.png"},{"id":63812312,"identity":"5c1b565f-01a8-42df-a1fc-111bacfe7872","added_by":"auto","created_at":"2024-09-02 14:11:05","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1077986,"visible":true,"origin":"","legend":"\u003cp\u003ePictures of physiological appeareance of Pleurotus ostreatus on different substrate combinations\u003c/p\u003e\n\u003cp\u003eA=cotton hulls and sawdust,b=cotton hulls and banana fronds,c=cotton hulls and wheatstraw, d=cotton hulls and maizecobs\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-4831136/v1/d0085fc2d3d3bf2f69d7b17f.png"},{"id":66776829,"identity":"c6e332df-feba-4d1e-b9c7-0d298d11c694","added_by":"auto","created_at":"2024-10-16 11:16:50","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2366012,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4831136/v1/5d95bdee-76f5-45fb-82d6-317cb14dbd05.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Evaluation of wheatstraw, sawdust, banana fronds, maize cobs and cotton hulls substrate combinations for Pleurotus ostreatus cultivation.","fulltext":[{"header":"Introduction","content":"\u003cp\u003e\u003cem\u003ePleurotus ostreatus(\u003c/em\u003eoyster) is a member of \u003cem\u003ePleurotus\u003c/em\u003e species of mushrooms cultivated globally for food. According to Besufekud \u003cstrong\u003eet al\u003c/strong\u003e (2019), oyster mushrooms are the second largest commercially produced mushrooms in the world. They possess a rare capability to convert lignocellulosic waste residues into edible fruiting bodies, rich in nutrients. Oyster mushrooms are reported by Ramsbottom (\u003cspan class=\"CitationRef\"\u003e1954\u003c/span\u003e) to contain high amounts of vitamins, particularly vitamin D and high protein contents because they contain enzymatic proteases that can breakdown complex macromolecules in agricultural wastes into individual amino acids. Of much importance is the absence of fatty acids in oyster mushrooms, which makes them ideal for persons willing to maintain a low fat diet. Furthermore, oyster mushrooms contain the appropriate ratios of potassium and sodium, which are essential in reducing heart disease.\u003c/p\u003e\n\u003cp\u003eIn Zimbabwe, mushroom cultivation is still at the grassroots level with most small-scale farmers producing enough for subsistence. Much knowledge is still lacking pertaining to the utilisation of substrate individually and in combination to optimise production and achieve commercial production leading to exportation.Govera (2020)\u003c/p\u003e\n\u003cp\u003eMushroom substrate is the material or substance, which provides food for the growing mycelium. Oyster mushrooms can grow on a wide range of substrates compared to other mushrooms. They can grow on all wood types including sawdust and paper pulp sludge. They also grow on all types of cereal straw such as maize straw, wheatstraw and rice straw. They also grow well on other types of agricultural substrates such as corncobs, banana fronds, maize stalks and leaves, savanna veld grass, sage grass, cottonseed hulls and coffee grounds. The mechanism by which substrate is broken down, is by the extracellular release of enzymes.According to Onyeka et al (\u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e) the extracellular enzymes secreted by fungi contain amorphous homo and heteropolysaccharides often associated with fungal protein. These enzymes bind to cellulose, lignin and hemicellulose, and break them down into simple sugars and peptides.\u003c/p\u003e\n\u003cp\u003eSayner \u003cstrong\u003eet al\u003c/strong\u003e (2019) alludes that the yield and quality of oyster mushroom is largely dependent on the chemical and nutritional content of the substrate. In instances where, a nutrient lacking in one substrate it is then provided by another substrate. There is need to establish a consensus by, pulling high yielding substrates together and then come up with the best formulation.\u003c/p\u003e\n\u003cp\u003eMushroom yield is closely associated with the chemical and biological composition of the substrate (Bhattacharjya et al \u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e) Most mushroom farmers in Zimbabwe today are using cotton hulls to grow oyster mushroom. As more people are becoming knowledgeable about cotton hulls as substrate, it creates a demand for this commodity. Tapiwa (2021). As a result of the increase in demand for cotton hulls there is increase in price of cotton hulls. It becomes economically non profitable to venture into mushroom farming due to expensive substrate. The price of wheat straw and cotton hulls is increasing every year, as many more people become mushroom farmers. This in turn affects the pricing of the mushrooms themselves.\u003c/p\u003e\n\u003cp\u003eDead plant matter has different composition of nutrition. There are many factors, which determine selection of a particular substrate. These are nutrition, cost, availability, accessibility and storage. A single particular substrate may be readily available but costly to the farmer. The length of the mushroom crop or the number of flushes depends on the substrate used for the cultivation of oyster mushrooms. For example, wheatstraw has good nutrition but it has a limited cropping life. Banana fronds also have good nutrition but they has low water retention. Sawdust has high nutrition but it takes longer to fruit. Cotton hulls are the best for fruiting, spawn run, have good nutrition but they are very expensive and seasonal (Pant et al \u003cspan class=\"CitationRef\"\u003e2006\u003c/span\u003e). There are limitations in nutritional composition when only one substrate is used to grow mushrooms. A formulation of substrates might contribute to better nutrition in oyster mushrooms. The best composition of substrates should have a short spawn run, a high biological efficiency and a high benefit cost ratio. C/N ratio directly affects the mushroom spawn run, yield and biological efficiency.\u003c/p\u003e\n\u003ch3\u003eSpawn run\u003c/h3\u003e\n\u003cp\u003eSpawn run is the time it takes for the substrate to be fully covered by the mushroom mycelia. It is affected by qualities of the substrate and qualities of the spawn. For a shorter spawn run a substrate should have a high surface area and good nutrition. Banana leaves require 19 days for spawn run.(Asmamaw \u003cstrong\u003eet al\u003c/strong\u003e 2014).Sawdust takes 30 days for spawn run. (Hoa et al 2015).Cotton hulls takes 18 days for spawn run.(Muswati et al 2021). Wheatstraw takes 16 days for spawn run.(Girmay et al \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e). Corn cobs takes 40 days for spawn run. (Itelima \u003cspan class=\"CitationRef\"\u003e2011\u003c/span\u003e)\u003c/p\u003e\n\u003ch3\u003eBiological efficiency\u003c/h3\u003e\n\u003cp\u003eBiological efficiency is defined as the percentage measurement of harvested fresh mushrooms relative to the dry weight of substrate.(Chukwurah \u003cstrong\u003eet al\u003c/strong\u003e 2012). In theory a 100% biological efficiency is achieved when fresh mushrooms having a moisture content of 90% are havested from a substrate with a moisture content of 75% of which 25% of dry substrate gives us the mushrooms.\u003c/p\u003e\n\u003cp\u003eBE\u0026thinsp;=\u0026thinsp;DP/M\u003c/p\u003e\n\u003cp\u003eBE-Biological Efficiency\u003c/p\u003e\n\u003cp\u003eDP-mass of fresh mushrooms produced in kgs\u003c/p\u003e\n\u003cp\u003eM-initial dry mass of substrate in kgs\u003c/p\u003e\n\u003cp\u003eIn practice a 100% biological efficiency is achieved when a mushroom farmer collects 10 kgs of mushrooms from three harvests after cultivating on 10kgs of dry substrate. Outside factors such as temperature and humidity, affect biological efficiency. (Girmay 2016)\u003c/p\u003e\n\u003cp\u003eA biological efficiency above 75% will be economical for the mushroom farmer. This is only achievable by using a high spawn rate. Usually the first, second and third flushes are the best flushes, with each flush decreasing in yield form the first. For subsistence, small scale farmers can allow the mushroom bags to fruit until the substrate is spent. For commercial farming only three flushes are economical. (Zakil \u003cstrong\u003eet al\u003c/strong\u003e 2019)\u003c/p\u003e\n\u003ch3\u003eB-C Ratio\u003c/h3\u003e\n\u003cp\u003eB-C (Benefit-Cost) ratio is the ratio of benefit from selling output monetary terms relative to the total cost during production. However, as the value of B:C increases so does the feasibility of mushroom production in economical aspect. To find the ratio the yield of mushrooms by is divided by the the sum total expenditure of raw materials and substrate. Rice straw gives a B-C ratio of 3.498, wheatstraw 1.108, sugarcane bargasse 0.217 and banana leaves 1.408. Therefore, as the B-C ratio becomes greater then 1, the more profitable the project and as the B-C ratio falls below 1, the less profitable the project. (Dubey et al \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n\u003ch2\u003eCarbon to Nitrogen ratio\u003c/h2\u003e\n\u003cp\u003eAccording to Kashangura et al (\u003cspan class=\"CitationRef\"\u003e2005\u003c/span\u003e)for mycelial growth and fruiting body formation the carbon to nitrogen ratio should be 50: 1, 100: 1 or 500: 1. Most agricultural wastes such as corncob, grain straws, wood, sawdust and banana fronds have sufficient C; N ratios to support saprophytic mushroom growth. C: N ratio is the proportion of carbon to nitrogen in a substrate. (Hoa \u003cstrong\u003eet al\u003c/strong\u003e 2015)\u003c/p\u003e\n\u003cp\u003eOyster mushroom mycelial growth is optimum in the presence of more carbon compared to nitrogen. Therefore, substrates comprised primarily of cellulose, hemicellulose and lignin are good candidates for oyster mushroom cultivation. In this study, we will investigate cotton hulls, sawdust, wheatstraw, banana fronds and maize cobs individually and in combination to determine potential in optimisation of \u003cem\u003ePleurotus ostreatus\u003c/em\u003e. We discuss the characteristic features and C: N ratios of the substrates under investigation below:\u003c/p\u003e\n\u003cp\u003eBanana fronds are the dried leaves from banana plants. During the dry season banana leaves lose a lot of moisture and as a result, the plant withholds water in the stem, leading to the lower leaves turning brown and then drooping down the plant. Banana farmers have no choice but to cut away the leaves and burn them because the leaves invite pests such as mosquitoes, rats and fungi, which feed on the dead leaves. The carbon to nitrogen ratio in banana leaves is 70:1. (ScienceMate \u003cspan class=\"CitationRef\"\u003e2022\u003c/span\u003e)\u003c/p\u003e\n\u003cp\u003eWheatstraw is the grass or straw that farmers leave behind when wheat farmers harvest their wheat. Wheat is a winter crop with a very short season and farmers have to use combine harvesters to harvest the wheat before the pod breaks and disperses the seed. Most farmers use a bailer to collect the leftover straw and then store it for feeding their livestock. Wheatstraw has problems with storage since it easily burns and termites tend to feed on the wheat straw. Less knowledgeable farmers tend to burn the remaining wheatstraw left in the fields in preparation for the maize farming season in Zimbabwe. This however leads to veld fires and deforestation. The carbon to nitrogen ratio in wheat is 80:1. (Lentz and Lindsey 2016)\u003c/p\u003e\n\u003cp\u003eMaize cobs are the cob that remains after removal of maize seeds from the cob. Large and huge masses of this waste accumulates soon after maize has been harvested because fresh maize cobs are sold and cooked as breakfast and discarded along streets and at dumpsites creating a huge environmental problem. Most of the weight of the maize cob is in the cob itself and this proves that it has a lot of carbon. Grinding down the cob after drying it can drastically increase its surface area thereby allowing mushroom mycelia to spread and absorb nutrients from the cob. Postharvest, farmers discard the cobs after shelling and most small-scale farmers use them as firewood during the winter. The carbon to nitogen ratio of maize cobs is 100:1.Adjapong (2015)\u003c/p\u003e\n\u003cp\u003eThe wood industry is reponsible for producing furniture and roofing material. However, during the cutting down, shaping and designing a lot of this wood is broken down into wood shavings and sawdust. However if the wood waste derived from this industry was to be saved stored and commercialised for mushroom farming we could archive food security. The carbon to nitrogen ratio of wood is in the range of 500:1. (Rynk \u003cspan class=\"CitationRef\"\u003e1992\u003c/span\u003e)\u003c/p\u003e\n\u003cp\u003eCottonseed hulls are a waste product resulting from the pressing and crushing down of cottonseed into cottonseed cooking oil. Ginneries opt to burn or throw away the waste as the main method of disposal. However, cottonseed hulls have proved to contain high amounts of nitrogen and carbon essential for mushroom growth. The carbon to nitrogen ratio of cottonseed hulls is 59:1. (Haziran \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e)\u003c/p\u003e\n\u003cp\u003eThis study will provide sufficient data to make decisions on the utilisation of different types of agricultural and forest waste into nutritional food. To analyse, evaluate and ultimately identify the most appropriate composition of substrate for cultivating \u003cem\u003ePleurotus ostreatus\u003c/em\u003e.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eSite of the study\u003c/h2\u003e \u003cp\u003eThe research was conducted in a steel Mushroom House, at the Department of Biotechnology, School of Health Sciences and Technology, Chinhoyi University of Technology, Zimbabwe from January to March 2022 for evaluating the effect of different substrates on the perfomance of \u003cem\u003ePleurotus ostreatus\u003c/em\u003e.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eTreatments\u003c/h3\u003e\n\u003cp\u003eThe substrates used in this investigation were wheatstraw, cottonseed hulls, sawdust, banana fronds and maize cobs. Each substrate combination weighing 1kg (dry weight) at a ratio of 1:1 was also distributed into a single factor experiment laid out in a Completely Randomised Design Design. Nine treatments were replicated four times thus making 36 bags.\u003c/p\u003e \u003cp\u003eAnalysis of variance tested for differences among means and means separated using Least Significant Difference (LSD) at the 5% level of significance. One-way analysis of variance (ANOVA) with Duncan\u0026rsquo;s multiple range tested the mean significant differences (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) among treatments by using computer software R Statistical Package.\u003c/p\u003e \u003cp\u003eSubstrate collection and bag filling:\u003c/p\u003e \u003cp\u003eDifferent substrates were collected from different locations. Cotton hulls purchased from a retailer in Harare. Maize cobs purchased from Citrus farm in Chinhoyi. Banana fronds collected from Bhagudha farm Chinhoyi. Sawdust procured from Chinhoyi University Works and Estates Department. Wheatstraw was collected from Chinhoyi University farm Chinhoyi.\u003c/p\u003e \u003cp\u003eMaize cobs, wheatstraw and banana fronds were ground at the Chinhoyi University feed factory. Maize cobs were ground into powder, wheatstraw into chunks 2-4cm and banana fronds into chunks 2\u0026ndash;4 cm long. Combinations formulated with respect to cotton hulls at a ratio of 1:1 dry weight. They were soaked with clean tap water to remove all dust particles. Then the substrate combinations filled into separate 200litre drums and a mixture of 4% sodium hypochlorite and 20% cacium hydroxide solution added to cover the substrate. The drums were covered and left overnight. The following day, substrates were drained of excess water separately on steel fence tables. Each substrate was mixed with an equal proportion of cotton hulls.\u003c/p\u003e \u003cp\u003eInoculation/spawning:\u003c/p\u003e \u003cp\u003eWe made our own \u003cem\u003ePleurotus ostreatus\u003c/em\u003e grain spawn to avoid bias. Spawning was carried out in transparent polythene pastic bags in layers subsequently with the first layer consisting of substrate and the top layer covered with spawn. Each bag comprised of 3 layers of substrate and 3 layers of spawn. After inoculation each bag approximately measured 2kg of substrate combinations by wet weight. Then the mouth of each bag was secured tightly with string and small holes were punched on the top and lateral sides of each bag for aeration.\u003c/p\u003e \u003cp\u003eIncubation:\u003c/p\u003e \u003cp\u003eThe packed bags were incubated in a dark steel mushroom house until the mycelium penetrated to the bottom of the bag. The bags were hung from trusses welded below the roof of the mushroom house to allow excess water to drip from the bottom of the bag, to prevent direct contact of the bags with the floor of the mushroom house and to prevent contact with insects such as ants and termites.\u003c/p\u003e \u003cp\u003eGrowing:\u003c/p\u003e \u003cp\u003eFrom the moment bags were introduced into the mushroom house the spawn run was observed and measured until the whole bags were covered with white mycelium. Thereafter the plastics were removed from the bags and the bags were placed on a wooden platform 30cm from the floor with a space of 20cm inbetween bags. When pinheads started appearing a light source was introduced into the mushroom house at night and watering commenced 3 times a day at 6 hour intervals using a knapsack sprayer.\u003c/p\u003e \u003cp\u003eHarvesting:\u003c/p\u003e \u003cp\u003eAfter 5 days of watering mushrooms were harvested and appropriate observations and measurements were taken. Harvesting commenced when the cap attained the maximum diameter, just before the edges flatten out. Firstly the whole bag and mushroom weight was recorded, then picking was done by gently twisting and pulling out the stalk. Mushrooms were then weighed and packaged into 200g punnets, wrapped with transparent plastic wrapper and sold for \u003cspan\u003e$\u003c/span\u003e1/200g. Then the weight of the empty bag was recorded. The benefit from selling the mushrooms was also recorded.\u003c/p\u003e \u003cp\u003eData collection and analysis:\u003c/p\u003e \u003cp\u003eData was recorded periodically during the incubation from colonisation to final substrate weight. Parameteres under conisderation were number of days to full colonisation, weight of bag and mushroom, total mushroom yield, final substrate weight, total mushrooms sold and benefit cost ratio. Collected data were analysed using R statistical software and Microsoft Excel 2007.\u003c/p\u003e \u003cp\u003eAnalysis of variance (ANOVA) was used to test among treatments and means were separated using Least Significance Difference (LSD) at the 5% level of significace.\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003eThe yield and yield attributing observations of mushrooms obtained from different combinations of substrates were compared. There was notable and considerable variation with respect to spawn run, quantiy of first yield and final substrate weight.\u003c/p\u003e \u003cp\u003eEffect of substrate combinatins on spawn run and 1st yield weight:\u003c/p\u003e \u003cp\u003eHighly significant results were observed among treatments in terms of spawn run of \u003cem\u003ePleurotus ostreatus\u003c/em\u003e as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Of all the combinations under investigation wheatstraw and cotton hulls combination took the shortest time (12 days) followed by cotton hulls and banana fronds(13 days), cotton hulls and maizecobs(13 days) and lastly cotton hulls and sawdust(17 days). Individual substrate took longer to complete spawn run compared to combinations i.e wheatstraw(16 days), banana fronds(17 days), maize cobs (19 days), cotton hulls(19 days ) and finally sawdust(20 days). This coincides with the findings of Muswati \u003cb\u003eet al\u003c/b\u003e 2021 that combinations of cotton hulls perform much better during spawn run compared to individual substrates.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMean values of first yield, spawn run and final substrate\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSubstrate\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirst yield(g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSpawn run(days)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSubstrate final weight(g)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCotton hulls and wheatstraw\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e376\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1576\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCotton hulls and banana fronds\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e449\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1716\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCotton hulls and sawdust\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e428\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1099\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCotton hulls and maize cobs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e420\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1508\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCotton hulls\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1213\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWheatstraw\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e212\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e980\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSawdust\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e155\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1417\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBanana fronds\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1118\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMaizecobs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e263\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1842\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eAnova table for analysis of variance on substrate means\u003c/h2\u003e \u003cp\u003eDf Sum Sq Mean Sq F value Pr(\u0026gt;\u0026thinsp;F)\u003c/p\u003e \u003cp\u003etm 3 11217444 3739148 146.8\u0026thinsp;\u0026lt;\u0026thinsp;2e-16 ***\u003c/p\u003e \u003cp\u003eResiduals 32 815253 25477\u003c/p\u003e \u003cp\u003e---\u003c/p\u003e \u003cp\u003eSignif. codes: 0 \u0026lsquo;***\u0026rsquo; 0.001 \u0026lsquo;**\u0026rsquo; 0.01 \u0026lsquo;*\u0026rsquo; 0.05 \u0026lsquo;.\u0026rsquo; 0.1 \u0026lsquo; \u0026rsquo; 1\u003c/p\u003e \u003cp\u003eAt the 5% level of significance a P value of \u0026lt;\u0026thinsp;2e-16 is less than 0.05 which means there are significant differences among treatment means.\u003c/p\u003e \u003cp\u003e \u003cb\u003eEffect of different substrates on total yield,spawn run and final substrate weight of oyster mushroom. (\u003c/b\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cb\u003e).\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe weight of substrate combination after first harvest was lowest in case of cotton hulls and sawdust(1099g) followed by cottonhulls maizecobs, cottonhulls-wheatstraw and cottonhulls-banana fronds at 1508g, 1576g and 1716g respectively as shown in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. This is evidence that oyster mushroom absorbs nutrients more efficiently in substrates composed of cotton hulls and sawdust compared to other substrate combinations of cotton hulls. This might also give an indication of particle size and absorption efficiency given the small particle size of sawdust rendering nutrient accessibility easier. Thus, it also indicates that the high biological efficiency obtained from cottonhulls-sawdust combination (76%) is attributed to the small surface area. Statistical analysis of data generated showed that there are significant differences in the total yield of mushrooms obtained from different substrate combinations. Yields from individual substrates pale in comparison to substrate combinations with the exception of cotton hulls(400g) alone which exceeded combinations of cotton hulls and wheatstraw(376g). Combinations of substrates contribute to nutrient compensation as what is lacking in one substrate is supplemented from the other. The high biological efficiency exhibited by cotton hulls sawdust combination may be attributed to high lignolytic and cellulonitic activity of the substrates. Muswati \u003cb\u003eet al\u003c/b\u003e (2021)cultivated \u003cem\u003ePleurotus ostreatus\u003c/em\u003e on cotton hulls and baobab fruit shells and observed similar results.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eEconomic analysis(Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e)\u003c/h2\u003e \u003cp\u003eCost of cultivation:\u003c/p\u003e \u003cp\u003eIt is analysed based on the cost of different materials and combinations of substrate. However, in this research the cost of required materials was at a constant for each treatment, except for cost of substrates, which are shown in Table\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e:\u003c/p\u003e \u003cp\u003eB-C ratio:\u003c/p\u003e \u003cp\u003eThe B: C ratio relates the benefit from selling mushrooms in monetary terms to the total cost during production. In this case it was calculated in United States Dollars by dividing the cost of production per kilogram of substrate by the total benefit of selling fresh mushrooms per kilogram of substrate. Cost of producton entails total expenses of raw materials. Benefit of production is calculated by dividing the biological efficiency by the price of selling mushrooms per kilogram of substrate. Substrate combinations were considered profitable if the B: C ratio was greater than 1. The differences in B:C ratio are shown in the Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The highest most significant ratio was produced by combination of cotton hulls and sawdust(5.7) whereas the lowest least significant ratio was produced by combination of cotton hulls and wheatstraw. For individual substrates cotton hulls produced a significant ratio of 4.24 which was more profitable compared to combinations of cotton hulls with banana fronds (2.43), cotton hulls with wheatstraw (2.02) and cotton hulls with maizecobs(3.73). For individual substrates cotton hulls(4.24) were more profitable compared to all the other substrates with the least profitable being banana fronds with a B;C ratio of 1.28. The data generated here is empirical basis for encouraging mushroom farmers to consider combining cotton hulls and sawdust for achieving high economic return with low investment. We suggest the low yied from wheatstraw, maizecobs, sawdust and banana fronds emanating from their high water retenton tendencies in contrast to cotton hulls. Furthermore wheatstraw and banana fronds lack compactness due to the low surface area which impedes complete ramification of substrate. Combinations of cotton hulls with wheatstraw and banana fronds did not improve yield due to reasons previously stated. Sawdust and cotton hulls proved to be the ideal combination firstly as a result of the large surface area then secondly due to the rich ratios of lignin and cellulose from sawdust combined with nitrogen and xylanases provided by cotton hulls.(Vhargese and Amritkumar 2020)\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMean values of B:C ratio of individual substrates and combinations\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSubstrate\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBiological efficiency/kg\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCost/kg(US\u003cspan\u003e$\u003c/span\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eBenefit(US\u003cspan\u003e$\u003c/span\u003e)/kg\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eB:C ratio\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCotton hulls and wheatstraw\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCotton hulls and banana fronds\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e45.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.43\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCotton hulls and sawdust\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e76.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e5.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCotton hulls and maize cobs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.73\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCotton hulls\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e60.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.015\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e4.24\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWheatstraw\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e25.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.85\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSawdust\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.925\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.81\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBanana fronds\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMaizecobs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e26.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.315\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.31\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eSpawn run was fastest for wheatstraw(16days) due to high nutrient availability and aeration. As the degreee of aeration decreases as a result of compactnaess of the substrate there is increased longevity of the spawn run as observed for sawdust(20 days) as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.. Spawn runner was also faster for banana fronds (17 days) also due to availability of aeration. Maizecobs and cotton husks managed to achieve the same rate of spawn runner dur tocompactness. It was observed that irregardless of nutrient availability particle size of substrate directly affects availability of air pockets within the substrate thereby limiting aiflow and affecting rate of fermentation. The more compact a substrate is the lesser the airation results in a slow spawn run.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eBiological efficiency is directly affected by the carbonn to nitrogen ratio. The more nitrogen there is in a substrate the higher the rate of protein synthesis leading to a higher yield of mushrooms(cotton husks). In contrast the more carbon there is in a substrate the more the more enegy available for cellular replication(mitosis) but less energy available for protein sythesis,as in the case of wheatstraw and sawdust. However complentation comes into play when the shortfalls of one substrate is covered by suplementation with another substrate resulting in a high biological efficiency as in the case of cotton hulls and sawdust as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eSpawn run is fastest for cotton husk and wheatstaw combinations. Spawn run is slowest for cotton husks and sawdust. The imrovement in spawn run for cotton hulls from 19 days to a shorter time of 12 days is abbtributed to increased aeration. Furthermore there is a notable decrease in the spawn ru for sawdust from 20 days to 17 days attributed by increased carbon to nitrogen ratio and aeration. Overally combinations of substrate perfomed better compared to indiividual substrate across the board.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigure\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e is data translated from Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e giving a graphical representation of the benefit cost ratio of using combinations of substrated compared to individual substrate. The highest benefit being afforded by combining cotton hulls and sawdust and the lowest being cotton hulls and wheatstraw. The only exception being cotto husk individully outperforming combinations with maizecobs, wheatstraw and banana fronds.\u003c/p\u003e \u003cp\u003e \u003cp\u003eFigure\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e shows appearance of oyster mushrooms cultivated on different combinations of substrate. On cotton hulls and sawdust, oyster mushrooms are bigger and there is uniform distribution of bunches right round the bag, there is fruiting on all sides of the bag. On cotton hulls and banana fronds there are lesser bunches of mushrooms even though there is also an even distribution of mushrooms right round the bag. On cotton houlls and wheatstraw the mushrooms have a much thicker skin and appear more grey than other mushrooms grown on other combinations. However, there are fewer mushrooms only appearing on one side of the bag. On cotton hulls and maizecobs there is a uniform distribution of mushrooms right round the bag however, the mushrooms appear smaller than the ones on cotton hulls and sawdust. The mushrooms on cotton hulls and maizecobs combination are whiter than mushrooms grown on other mushrooms.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study was conducted by growing \u003cem\u003ePleurotus ostreatus\u003c/em\u003e on 5 different agricultural waste substrates namely cotton hulls, maizecobs, wheatstraw, banaba fronds, sawdust and on combinations of 4 substrates with respect to cotton hulls. Among the individual substrates wheatstraw colonised the fastest whereas among the combinations wheatstraw and cotton hullls were colonised fastest. In terms of biological efficiency, individually cotton hulls performed exceedingly well compared to other substrates. However, combinations of substrates revealed a combination of sawdust and cotton hulls to be superior to other combinations. Combinations of substrates proved to be highly efficient both in spawn run and biological efficiency compared individual substrates. In terms of economic return, combinations of sawdust and cottonhulls (5.7) proved to be the most profitable. Hence, we conclude that in terms of spawn run, yield and economic return combination of cotton hulls and sawdust is highly recommended for farmers involved in the cultivation of \u003cem\u003ePleurotus ostreatus\u003c/em\u003e.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003e\u003cu\u003eEthical approval\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSince no humans or animals were involved in this study no ethical committees were consulted.\u003c/p\u003e\n\u003cp\u003eConsent to participate is not applicable since no human or animal participants were required.\u003c/p\u003e\n\u003cp\u003eAll supervisors and fellow co-authors have provided their consent to publish.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eFunding\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was self funded by the corresponding author Elyson Silivani as an Mphil Student in the School of Health Sciences, Department of Biotechnology, Chinhoyi University of Technology.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eHiMr Elyson Silivani and Dr Chigu wrote the main manuscriptDr Chisango and Professor Chitindingu reviewed the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBesufekad Yohannes, Mekonnen Abraham, Girma Bikila, Daniel Robel, Getahun Tassema, Melkamu Jale, Asefa, Malesu Fikiru Tsehaynesh, Denboba Lalise,(2019) \u003cem\u003eSelection of appropriate substrate for production of oyster mushroom\u003c/em\u003e (\u003cem\u003ePleurotus ostreatus\u003c/em\u003e)\u0026hellip; College of Natural and Computational Science, Department of Biotechnology, Wolkite University, Ethiopia.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRamsbottom J,(1954),\u003cem\u003eMushrooms and Toadstools:A study of the activities of fungi.\u003c/em\u003eCollins,London.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGovera C.,EcoAfrica,(2020), Chido Govera teaches young Zimbabwean women the art of Mushroom farming, Availableonlineat \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://m.youtube.com/watch?v=nDud2g1RVY\u003c/span\u003e\u003cspan address=\"https://m.youtube.com/watch?v=nDud2g1RVY\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e accessed 15/06/2020\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOnyeka, E. 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Fr.) Kumm (oyster mushroom) on different substrates\u003c/em\u003e. AMB Expr 6, 87 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s13568-016-0265-1\u003c/span\u003e\u003cspan address=\"10.1186/s13568-016-0265-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMuswati Charles, Simango Kennedy, Tapfumaneyi Linda, Mutetwa Moses, Ngezimana Wonder,(2021) \u003cem\u003eThe Effects of Different Substrate Combinations on Growth and Yield of Oyster Mushroom (Pleurotus ostreatus)\u003c/em\u003e Hindawi International Journal of Agronomy Volume 2021, Article ID 9962285, Available online at \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1155/2021/9962285\u003c/span\u003e\u003cspan address=\"10.1155/2021/9962285\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVarghese Bably Pearl, Amritkumar Pavithra,(2020) Comparative Study on Cultivation of Oyster Mushrooms using Nutrition Enhancing Substrates, International Journal of Scientific Research in Biological Sciences Vol.7, Issue.2, pp.105\u0026ndash;111, April (2020) E-ISSN: 2347-7520 DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.26438/ijsrbs/v7i2.105111\u003c/span\u003e\u003cspan address=\"10.26438/ijsrbs/v7i2.105111\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eItelima J. U.,(2011), Cultivation of mushroom (Pleurotus ostreatus) using corn cobs and saw dust as the major substrates, Global journal of agricultural sciences vol 11, no. 1, 2012: 51\u0026ndash;56 DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.4314/gjass.v11i1.9\u003c/span\u003e\u003cspan address=\"10.4314/gjass.v11i1.9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Substrate, combination, mushrooms, Pleurotus ostreatus, biological efficiency, spawn run, cost-benefit ratio","lastPublishedDoi":"10.21203/rs.3.rs-4831136/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4831136/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe yield and quality of oyster mushroom is largely dependent on the chemical and nutritional content of the substrate. The objective of the study was to determine the effect of different individual substrates and substrate combinations on the perfomance of \u003cem\u003eP.ostreatus\u003c/em\u003e for growth and sustainable development of the mushroom cultivation industry in Zimbabwe. Various individual substrates used as treatments, selected for the cultivation of oyster mushrooms were, wheatstraw, cotton hulls, sawdust, maize cobs and banana fronds each of 1kg and replicated 4 times. Various substrate combinations, also used as treatments were, cotton hulls and wheatstraw, cotton hulls and sawdust, cotton hulls and maizecobs, cotton hulls and banana fronds in the ratio 1:1. Each combination weighed 1kg and was replicated 4 times.Compound substrates perfomed much better in terms of biological efficiency and spawn run compared to individual substrates. The highest biological efficiency (76%) and spawn run(17 days) were obtained from combining cotton hulls and sawdust. Spawn run was fastest(12 days) for cotton hulls and wheatstraw combination. Analysis in terms of economic return, revealed that mushroom production was most profitable using cotton hulls and sawdust as substrate with a benefit-cost ratio of 5.7 compared to other combinations of agricultural residues. In terms of spawn run, yield and economic return, combining cotton hulls and sawdust is highly recommended for farmers involved in the cultivation of \u003cem\u003ePleurotus ostreatus\u003c/em\u003e to satisfy increasing consumer demand, whilst decreasing pressure on limited natural resources and preventing ecosystems degradation.\u003c/p\u003e","manuscriptTitle":"Evaluation of wheatstraw, sawdust, banana fronds, maize cobs and cotton hulls substrate combinations for Pleurotus ostreatus cultivation.","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-09-02 14:10:56","doi":"10.21203/rs.3.rs-4831136/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"807ac9b9-0b86-4532-b983-d07bfd93fadc","owner":[],"postedDate":"September 2nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-10-16T11:08:27+00:00","versionOfRecord":[],"versionCreatedAt":"2024-09-02 14:10:56","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4831136","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4831136","identity":"rs-4831136","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}